LOCUS Exported 7469 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION yeast plasmid with Hsp82. ACCESSION . VERSION . KEYWORDS p414CYC_Hsp82_CycTER SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7469) AUTHORS Jiang L, Mishra P, Hietpas RT, Zeldovich KB, Bolon DN TITLE Latent effects of Hsp90 mutants revealed at reduced expression levels. JOURNAL PLoS Genet. 2013 Jun;9(6):e1003600. doi: 10.1371/journal.pgen.1003600. Epub 2013 Jun 27. PUBMED 23825969 REFERENCE 2 (bases 1 to 7469) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7469 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(10..28) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_feature 70..573 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 610..714 /gene="bla" /label=AmpR promoter CDS 715..1575 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(933..952) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1746..2334 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2235..2254 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2488..2505 /label=L4440 /note="L4440 vector, forward primer" promoter 2658..2688 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2696..2712 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2701..2723 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2720..2736 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2720..2736 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2755..2775 /label=T3 /note="T3 promoter, forward primer" promoter 2757..2775 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 3104..3121 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(5300..5316) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5301..5317) /label=pBluescriptKS /note="For pBluescript vector" terminator 5319..5566 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(5335..5353) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(5584..5603) /label=T7 /note="T7 promoter, forward primer" promoter complement(5585..5603) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5613..5630) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5613..5629) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5622..5644) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 5774..6229 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5861..5880) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6071..6092 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(6328..7002) /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(7003..7283) /gene="S. cerevisiae TRP1" /label=TRP1 promoter primer_bind complement(7300..7319) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7419..7441 /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata 121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt 181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa 241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata 301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct 361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata 421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact 481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa 541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg 601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc 2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac 2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact 2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg 2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt 2761 aaccctcact aaagggaaca aaagctggag ctcatttggc gagcgttggt tggtggatca 2821 agcccacgcg taggcaatcc tcgagcagat ccgccaggcg tgtatatata gcgtggatgg 2881 ccaggcaact ttagtgctga cacatacagg catatatata tgtgtgcgac gacacatgat 2941 catatggcat gcatgtgctc tgtatgtata taaaactctt gttttcttct tttctctaaa 3001 tattctttcc ttatacatta ggacctttgc agcataaatt actatacttc tatagacacg 3061 caaacacaaa tacacacact aatctagaac tagtatgggt ggtcaccacc atcaccacca 3121 cggtggtcat atggctagtg aaacttttga atttcaagct gaaattactc agttgatgag 3181 tttgatcatc aacaccgtct attctaacaa ggaaattttc ttgagagaac tgatatctaa 3241 tgcctcggat gcgttggata aaattagata caaatctttg tctgatccaa agcaattgga 3301 aacagaacca gatctcttta ttagaatcac tccaaagcca gagcaaaaag ttttggaaat 3361 cagagattct ggtattggta tgaccaaggc tgaattgatt aataacttgg gtaccattgc 3421 caagtctggt accaaagcct tcatggaagc tctatctgct ggtgccgatg tatccatgat 3481 tggtcaattc ggtgttggtt tttactcttt attcttagtt gccgacagag ttcaggttat 3541 ttcaaagagc aacgacgacg aacaatacat ctgggaatcc aacgctggtg gttctttcac 3601 tgttactcta gacgaagtta atgaaagaat tggtaggggt accatcttga ggttattctt 3661 gaaagatgac caattggagt acttggaaga aaagagaata aaggaagtta tcaagagaca 3721 ttctgagttc gtggcctacc caatccaatt agtcgtcacc aaggaagttg aaaaggaagt 3781 tccaattcca gaagaagaaa agaaagacga ggaaaagaag gatgaggaaa agaaggatga 3841 agacgacaag aaaccaaaac tcgaggaagt cgatgaagaa gaggaaaaga agccaaagac 3901 gaaaaaagtt aaagaagaag ttcaagagat agaagaacta aacaagacta agcctttgtg 3961 gactagaaac ccatctgata tcactcaaga agaatacaat gctttctata agtctatttc 4021 aaacgactgg gaagacccat tgtacgttaa gcatttctcc gttgaaggtc aattggaatt 4081 tagagctatc ttattcattc caaagagagc accattcgac ttgtttgaga gtaaaaagaa 4141 gaagaataat atcaagttgt acgttcgtcg tgttttcatc actgatgaag ctgaagactt 4201 gattccagag tggttatctt tcgtcaaggg tgttgttgac tctgaggatt taccattgaa 4261 tttgtccaga gaaatgttac aacaaaataa gatcatgaag gttattagaa agaacattgt 4321 caaaaagttg attgaagcct tcaacgaaat tgctgaagac tctgaacaat ttgaaaagtt 4381 ctactcggct ttctccaaaa atatcaagtt gggtgtacat gaagataccc aaaacagggc 4441 tgctttggct aagttgttac gttacaactc taccaagtcc gtagatgagt tgacttcctt 4501 aactgattac gttaccagaa tgccagaaca ccaaaagaac atctactaca tcactggtga 4561 atctctaaag gctgtcgaaa agtctccatt tttggatgcc ttgaaggcta aaaacttcga 4621 ggttttgttc ttgaccgacc caattgatga atacgccttc actcaattga aggaattcga 4681 aggtaaaact ttggttgaca ttactaaaga tttcgaattg gaagaaactg acgaagaaaa 4741 agctgaaaga gagaaggaga tcaaagaata tgaaccattg accaaagctt tgaaagaaat 4801 tttgggtgac caagtggaga aagttgttgt ttcttacaaa ttgttggatg ccccagctgc 4861 tatcagaact ggtcaatttg gttggtctgc taacatggaa agaatcatga aggctcaagc 4921 cttgagagac tcttccatgt cctcctacat gtcttccaag aagactttcg aaatttctcc 4981 aaaatctcca attatcaagg aattgaaaaa gagagttgac gaaggtggtg ctcaagacaa 5041 gactgtcaag gacttgacta agttattata tgaaactgct ttgttgactt ccggcttcag 5101 tttggacgaa ccaacttcct ttgcatcaag aattaacaga ttgatctctt taggcctgaa 5161 cattgatgag gatgaagaaa cagagactgc tccagaagca tccaccgcag ctccggttga 5221 agaggttcca gctgacaccg aaatggaaga ggtagattaa tggatccccc gggctgcagg 5281 aattcgatat caagcttatc gataccgtcg acctcgagtc atgtaattag ttatgtcacg 5341 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc 5401 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat 5461 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa 5521 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat 5581 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac 5641 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 5701 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 5761 ggcgaatggc gcgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 5821 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 5881 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 5941 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 6001 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 6061 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 6121 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 6181 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc ctgatgcggt 6241 attttctcct tacgcatctg tgcggtattt cacaccgcat aggcaagtgc acaaacaata 6301 cttaaataaa tactactcag taataaccta tttcttagca tttttgacga aatttgctat 6361 tttgttagag tcttttacac catttgtctc cacacctccg cttacatcaa caccaataac 6421 gccatttaat ctaagcgcat caccaacatt ttctggcgtc agtccaccag ctaacataaa 6481 atgtaagctt tcggggctct cttgccttcc aacccagtca gaaatcgagt tccaatccaa 6541 aagttcacct gtcccacctg cttctgaatc aaacaaggga ataaacgaat gaggtttctg 6601 tgaagctgca ctgagtagta tgttgcagtc ttttggaaat acgagtcttt taataactgg 6661 caaaccgagg aactcttggt attcttgcca cgactcatct ccatgcagtt ggacgatatc 6721 aatgccgtaa tcattgacca gagccaaaac atcctcctta ggttgattac gaaacacgcc 6781 aaccaagtat ttcggagtgc ctgaactatt tttatatgct tttacaagac ttgaaatttt 6841 ccttgcaata accgggtcaa ttgttctctt tctattgggc acacatataa tacccagcaa 6901 gtcagcatcg gaatctagag cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt 6961 caccaatgga ccagaactac ctgtgaaatt aataacagac atactccaag ctgcctttgt 7021 gtgcttaatc acgtatactc acgtgctcaa tagtcaccaa tgccctccct cttggccctc 7081 tccttttctt ttttcgaccg aattaattct taatcggcaa aaaaagaaaa gctccggatc 7141 aagattgtac gtaaggtgac aagctatttt tcaataaaga atatcttcca ctactgccat 7201 ctggcgtcat aactgcaaag tacacatata ttacgatgct gtctattaaa tgcttcctat 7261 attatatata tagtaatgtc gtttatggtg cactctcagt acaatctgct ctgatgccgc 7321 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 7381 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 7441 gttttcaccg tcatcaccga aacgcgcga //